阅读说明：本技术 一种实时超声刺激电信号记录芯片及其制备方法 (Real-time ultrasonic stimulation electric signal recording chip and preparation method thereof ) 是由 孟龙 周伟 郑海荣 牛丽丽 林争荣 彭本贤 乌玛.儒可 崔英健 于 2020-12-10 设计创作，主要内容包括：本发明公开了一种实时超声刺激电信号记录芯片及其制备方法,该制备方法包括以下步骤：S1,在压电基底上制作叉指电极,得到声表面波芯片；并制作记录电极和电极引线S2,在S1得到的芯片上制作绝缘保护层,并对绝缘保护层处理形成记录电极,得到将叉指电极和记录电极结合的芯片；S3,制备PDMS腔道；S4,将S3制备的PDMS腔道与S2得到的芯片键合,得到实时超声刺激电信号记录芯片。本发明通过将产生声表面波超声的叉指电极与多通道记录电极结合,实现了超声刺激下多通道电信号的实时记录,能够实现神经环路上多个神经元电信号的实时记录,有效解决了传统换能器由于体积过大,与多通道记录电极不兼容的问题。(The invention discloses a real-time ultrasonic stimulation electric signal recording chip and a preparation method thereof, wherein the preparation method comprises the following steps: s1, manufacturing interdigital electrodes on the piezoelectric substrate to obtain a surface acoustic wave chip; manufacturing a recording electrode and an electrode lead S2, manufacturing an insulating protective layer on the chip obtained in S1, and processing the insulating protective layer to form the recording electrode, so as to obtain a chip combining the interdigital electrode and the recording electrode; s3, preparing a PDMS cavity channel; and S4, bonding the PDMS cavity channel prepared in the S3 with the chip obtained in the S2 to obtain a real-time ultrasonic stimulation electrical signal recording chip. The interdigital electrode generating surface acoustic wave ultrasound is combined with the multi-channel recording electrode, so that real-time recording of multi-channel electric signals under ultrasonic stimulation is realized, real-time recording of a plurality of neuron electric signals on a neural loop can be realized, and the problem that the traditional transducer is incompatible with the multi-channel recording electrode due to overlarge volume is effectively solved.)

1. A preparation method of a real-time ultrasonic stimulation electric signal recording chip is characterized by comprising the following steps:

s1, manufacturing interdigital electrodes on the piezoelectric substrate to obtain a surface acoustic wave chip, and manufacturing recording electrodes and electrode leads;

s2, manufacturing an insulating protection layer on the chip obtained in the S1, and processing the insulating protection layer to form a recording electrode to obtain a chip combining the interdigital electrode and the recording electrode;

s3, preparing a PDMS cavity channel;

and S4, bonding the PDMS cavity channel prepared in the S3 with the chip obtained in the S2 to obtain a real-time ultrasonic stimulation electric signal recording chip.

2. The method for preparing a real-time ultrasonic stimulation electrical signal recording chip according to claim 1, wherein in S1, an interdigital electrode is fabricated on a piezoelectric substrate to obtain a surface acoustic wave chip, specifically:

patterning the photoresist on the piezoelectric substrate through photoetching, forming a metal layer on the patterned piezoelectric substrate through magnetron sputtering, and removing the photoresist to obtain the surface acoustic wave chip.

3. The method for preparing a real-time ultrasound-stimulated electrical signal recording chip according to claim 2, wherein the step S1 is specifically as follows:

s11, gluing: spin-coating photoresist with the thickness of 1-5 μm on the surface of the piezoelectric substrate, and heating;

s12, exposure and development: covering the prepared film on the photoresist for exposure, and then developing by adopting a developing solution to pattern the photoresist;

s13, sputtering: carrying out magnetron sputtering on the patterned piezoelectric substrate to form a metal layer, and obtaining the piezoelectric substrate of the growth electrode;

s14, removing the photoresist: and (4) carrying out ultrasonic cleaning on the piezoelectric substrate of the growth electrode obtained in the step S13 in an acetone solution, and stripping the photoresist to obtain the surface acoustic wave chip.

4. The method for preparing a real-time ultrasound-stimulated electrical-signal recording chip according to claim 3, wherein the photoresist in S11 is a positive photoresist AZ5214, and the spin-coating conditions are as follows: rotating speed of 3000rpm for 30 s;

the film in the S12 is a film, and the developing solution is mif 300.

5. The method for preparing a real-time ultrasound-stimulated electrical signal recording chip according to claim 1, wherein the step S2 specifically comprises:

s21, sputtering: manufacturing an insulating protective layer on the surface acoustic wave chip obtained in the step S1 by a sputtering method;

s22, processing: and spin-coating photoresist on the insulating protective layer, covering the prepared film on the photoresist for exposure, developing by adopting a developing solution, and etching to form a recording electrode, thereby obtaining a chip combining the interdigital electrode and the recording electrode.

6. The method for preparing a real-time ultrasound stimulation electrical signal recording chip according to claim 5, wherein the photoresist in S22 is a negative photoresist SUN 1300; the spin coating conditions are as follows: rotating speed of 3000rpm for 30 s; the film is a film.

7. The method for preparing a real-time ultrasound-stimulated electrical signal recording chip according to claim 1, wherein the step of S3 preparing the PDMS cavity comprises:

manufacturing a PDMS cavity mold, casting PDMS in the PDMS cavity mold, and stripping after the PDMS is cured to obtain a PDMS cavity;

the PDMS cavity mold is manufactured by a three-dimensional printing method.

8. The method for preparing a real-time ultrasound-stimulated electrical-signal recording chip according to claim 1, wherein the bonding of the PDMS cavity and the chip in S4 comprises:

carrying out plasma treatment on the PDMS cavity and the chip, then bonding the PDMS cavity and the chip, and baking the PDMS cavity and the chip at 80 ℃ for 20min to obtain a real-time ultrasonic stimulation electric signal recording chip;

wherein the plasma treatment conditions are as follows: power 150W, time 70 s.

9. The method for preparing a real-time ultrasound-stimulated electrical signal recording chip according to any one of claims 1 to 8, wherein the piezoelectric substrate in S1 is a 128 ° YX double-side polished lithium niobate substrate, a zinc oxide substrate or an aluminum nitride substrate.

10. The real-time ultrasonic stimulation electric signal recording chip prepared by the preparation method according to any one of claims 1 to 9.

Technical Field

The invention belongs to the technical field of nerve regulation and control, and particularly relates to a real-time ultrasonic stimulation electric signal recording chip and a preparation method thereof.

Background

For a long time, mental diseases such as dyskinesia, pain, epilepsy, Parkinson's disease, mental diseases and angina pectoris seriously affect the health and quality of life of human beings. Despite the continuous clinical use of new antipsychotic drugs, a significant proportion of patients remain insensitive to medication or unsatisfactory in therapeutic efficacy. The nerve regulation and control treatment method belongs to a more popular treatment method in recent years, has a good treatment effect on various nerve diseases, and has a high development speed.

The nerve regulation is a biomedical engineering technology which plays a role in exciting, inhibiting or regulating neurons or nerve signal transduction at adjacent or distant parts of a central nervous system, a peripheral nervous system and an autonomic nervous system through an implantable or non-implantable technology and an electric or chemical action mode so as to achieve the aims of improving the life quality of patients and improving the nerve function.

Ultrasonic neuromodulation is a neuromodulation means proposed in recent years, which can non-invasively penetrate through a skull to regulate and control a nerve nucleus of a brain, and has attracted wide attention in the aspect of treating diseases such as Parkinson, epilepsy and the like, but the mechanism of ultrasonic neuromodulation is not clear. The nerve signal is an electric signal and shows potential change when the signal is transmitted on the nerve, so that the record of the electric signal can effectively represent the response of the neuron under the action of ultrasound so as to research the mechanism of ultrasound nerve regulation and control. However, the current commercial ultrasonic transducer is huge in volume, is difficult to be compatible with electrophysiological means, such as calcium imaging, patch clamp and other electric signal recording means, and the sound field of the traditional ultrasonic transducer is not uniform, and the ultrasonic stimulation energy to the recorded neuron depends heavily on the relative position of the recorded neuron and the transducer, so that the result is difficult to repeat. Therefore, it is necessary to develop a chip which has uniform ultrasonic energy and easy operation and can realize real-time recording of neuron electrical signals under ultrasonic stimulation.

Disclosure of Invention

In view of the above, the present invention provides a method for preparing a real-time ultrasound stimulation electrical signal recording chip, which realizes real-time recording of an electrical signal under ultrasound stimulation by bonding an interdigital electrode and a recording electrode to a PDMS channel.

The invention also aims to provide the real-time ultrasonic stimulation electric signal recording chip prepared by the preparation method.

The technical scheme adopted by the invention is as follows:

a method for preparing a real-time ultrasonic stimulation electric signal recording chip comprises the following steps:

s1, manufacturing interdigital electrodes on the piezoelectric substrate to obtain a surface acoustic wave chip, and manufacturing recording electrodes and electrode leads;

s2, manufacturing an insulating protection layer on the chip obtained in the S1, and processing the insulating protection layer to form a recording electrode to obtain a chip combining the interdigital electrode and the recording electrode;

s3, preparing a PDMS cavity channel;

and S4, bonding the PDMS cavity channel prepared in the S3 with the chip obtained in the S2 to obtain a real-time ultrasonic stimulation electric signal recording chip.

Preferably, in S1, an interdigital electrode is fabricated on a piezoelectric substrate to obtain a surface acoustic wave chip, specifically:

patterning the photoresist on the piezoelectric substrate through photoetching, forming a metal layer on the patterned piezoelectric substrate through magnetron sputtering, and removing the photoresist to obtain the surface acoustic wave chip.

Preferably, the S1 is specifically:

s11, gluing: spin-coating photoresist with the thickness of 1-5 μm on the surface of the piezoelectric substrate, and heating;

s12, exposure and development: covering the prepared film on the photoresist for exposure, and then developing by adopting a developing solution to pattern the photoresist;

s13, sputtering: carrying out magnetron sputtering on the patterned piezoelectric substrate to form a metal layer, and obtaining the piezoelectric substrate of the growth electrode;

s14, removing the photoresist: and (4) carrying out ultrasonic cleaning on the piezoelectric substrate of the growth electrode obtained in the step S13 in an acetone solution, and stripping the photoresist to obtain the surface acoustic wave chip.

Preferably, the photoresist in S11 is a positive photoresist AZ5214, and the spin coating conditions are as follows: rotating speed of 3000rpm for 30 s;

the film in the S12 is a film, and the developing solution is mif 300.

Preferably, the S2 is specifically:

s21, sputtering: manufacturing an insulating protective layer on the surface acoustic wave chip obtained in the step S1 by a sputtering method;

s22, processing: and spin-coating photoresist on the insulating protective layer, covering the prepared film on the photoresist for exposure, developing by adopting a developing solution, and etching to form a recording electrode, thereby obtaining a chip combining the interdigital electrode and the recording electrode.

Preferably, the photoresist in S22 is a negative photoresist SUN 1300; the spin coating conditions are as follows: rotating speed of 3000rpm for 30 s; the film is a film.

In specific implementation, the insulating protection layer is selected from at least one of silicon dioxide, photoresist, polyimide and silicon nitride, that is, any one of the above materials is used for preparing the insulating protection layer.

Preferably, the S3 is to prepare a PDMS channel, specifically:

manufacturing a PDMS cavity mold, casting PDMS in the PDMS cavity mold, and stripping after the PDMS is cured to obtain a PDMS cavity;

the PDMS cavity mold is manufactured by a three-dimensional printing method.

Preferably, the specific process of bonding the PDMS channel and the chip in S4 is as follows:

carrying out plasma treatment on the PDMS cavity and the chip, then bonding the PDMS cavity and the chip, and baking the PDMS cavity and the chip at 80 ℃ for 20min to obtain a real-time ultrasonic stimulation electric signal recording chip;

wherein the plasma treatment conditions are as follows: power 150W, time 70 s.

Preferably, the piezoelectric substrate in S1 is a 128 ° YX double-side polished lithium niobate substrate, a zinc oxide substrate, or an aluminum nitride substrate.

The invention also protects the real-time ultrasonic stimulation electric signal recording chip prepared by the preparation method.

The invention has the beneficial effects that:

1. the interdigital electrode is combined with the recording electrode, namely the interdigital electrode generating surface acoustic wave ultrasound is combined with the multi-channel recording electrode, so that the real-time recording of multi-channel electric signals under ultrasonic stimulation is realized, the real-time recording of a plurality of neuron electric signals on a neural loop can be realized, and an ultrasonic neural regulation and control mechanism is researched on the neural loop; compared with a commercial multi-channel recording electrode, the size of the chip is not increased, so that the problems that the traditional transducer is too large in volume and is incompatible with the multi-channel recording electrode are effectively solved;

2. the relative position of the interdigital electrode and the neuron cell at the position of the recording electrode in the chip is fixed, so that the energy borne by the cell is the same under the same parameter, the repeatability of the experiment is ensured, and the accuracy of neuron electric signal recording is ensured;

3. through the setting of PDMS chamber, can place neuron cell or brain piece and cultivate in PDMS chamber, can realize the record of the supersound stimulation of long-term and the signal of telecommunication, can study the effect of the long-time effect of supersound.

Drawings

Fig. 1 is a diagram illustrating a process of manufacturing a surface acoustic wave chip in a method for manufacturing a real-time ultrasound-stimulated electrical signal recording chip according to embodiment 1 of the present invention;

in the figure: (a) the method comprises the steps of gluing a piezoelectric substrate; (b) exposure and development processes; (c) a sputtering process is adopted; (d) the process of obtaining the surface acoustic wave chip after removing the photoresist;

fig. 2 is a diagram of a process of manufacturing a chip in which an interdigital electrode and a recording electrode are combined in a method for manufacturing a real-time ultrasound-stimulated electrical signal recording chip according to embodiment 1 of the present invention;

in the figure: (a) a process of sputtering an insulating protective layer; (b) the processes of gluing, exposing and developing are carried out; (c) an etching process is carried out;

fig. 3 is a diagram of a preparation process of bonding a PDMS cavity and a chip in the preparation method of the real-time ultrasound stimulation electrical signal recording chip according to the invention 1.

In the figure: 1. a piezoelectric substrate; 2. photoresist; 3. the combination of the interdigital electrode and the recording electrode; 4. an insulating protective layer; 5. an etched insulation protection layer; 6. PDMS channels.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Example 1

The embodiment provides a method for manufacturing a real-time ultrasonic stimulation electrical signal recording chip, as shown in fig. 1 to 3, comprising the following steps:

s1, manufacturing interdigital electrodes on the piezoelectric substrate 1 to obtain a surface acoustic wave chip, and manufacturing recording electrodes and electrode leads;

the method specifically comprises the following steps:

s11, gluing: spin-coating positive photoresist AZ5214 on the surface of the completely and clearly cleaned piezoelectric substrate 1 at 3000rpm for 30s to obtain photoresist 2 with the thickness of 1-5 μm shown in FIG. 1(a), and baking on a heating plate at 65 ℃ for 3 min;

wherein the piezoelectric substrate 1 is a 128-degree YX double-side polished lithium niobate substrate;

s12, exposure and development: covering the prepared film on the photoresist 2 shown in the figure 1(a) for exposure, wherein the pattern part is opaque, the non-pattern part is transparent, and the part with light transmission is solidified; when the mif300 is adopted for development, the solidified part is dissolved, the non-solidified part is not dissolved, and the pattern shown in the figure 1(b) is formed by development;

s13, sputtering: performing magnetron sputtering on the patterned piezoelectric substrate 1 to form a metal layer with the thickness of 200nm, as shown in fig. 1(c), so as to obtain a piezoelectric substrate for growing electrodes;

s14, removing the photoresist: and (d) ultrasonically cleaning the piezoelectric substrate of the growth electrode obtained in the step S13 in an acetone solution, and stripping the photoresist to obtain the surface acoustic wave chip as shown in a figure 1 (d).

S2, forming an insulating protection layer 4, i.e. a silicon dioxide layer, on the chip obtained in S1, and processing the insulating protection layer 4 to form a recording electrode, as shown in fig. 2, to obtain a chip combining the interdigital electrode and the recording electrode, i.e. a combination 3 of the interdigital electrode and the recording electrode;

the method specifically comprises the following steps:

s21, sputtering: cleaning the prepared chip, and preparing an insulating protection layer 4 by a sputtering method, as shown in fig. 2 (a);

s22, processing: on the surface of a completely cleaned chip, spin-coating a negative photoresist SUN1300 at 3000rpm for 30s, then covering the prepared film sheet on the surface for exposure, wherein a pattern part is opaque, a non-pattern part is transparent, a part with light transmission is solidified, the solidified part is not dissolved when developing, the non-solidified part is not dissolved, and through the design of the film sheet, a solvent part is a recording electrode position, as shown in fig. 2 (b); etching the chip surface insulating protective layer by an etching method, etching the position protective layer without the photoresist to obtain an etched insulating protective layer 5, and displaying a recording electrode for recording an electric signal, as shown in fig. 2 (c);

s3, preparing a PDMS cavity channel 6;

the method specifically comprises the following steps:

s31, printing a PDMS cavity mold through 3D;

s32, casting PDMS: mixing the glue A and the glue B of the PDMS according to the mass ratio of 10:1, uniformly mixing, pouring into a mold, vacuumizing to remove bubbles in the PDMS, and finally placing the culture dish in an oven at 80 ℃ for 30min to solidify the PDMS;

s33, stripping: stripping the PDMS cavity channel 6;

and S4, bonding the PDMS cavity channel 6 prepared in the S3 with the chip obtained in the S2 to obtain a real-time ultrasonic stimulation electric signal recording chip.

The method specifically comprises the following steps: and carrying out plasma treatment on the chip manufactured in the step S2 and the PDMS cavity 6 manufactured in the step S3, wherein the power of the plasma treatment is 150W, the duration is 70S, then, adhering the PDMS cavity 6 on the chip for bonding, and baking in an oven at 80 ℃ for 20min to obtain the manufactured real-time ultrasonic stimulation electrical signal recording chip for the experiment shown in the figure 3.

The embodiment also provides the real-time ultrasonic stimulation electric signal recording chip prepared by the preparation method.

Example 2

The real-time ultrasound stimulation electrical signal recording chip and the manufacturing method thereof in this embodiment are the same as those in embodiment 1, except that the piezoelectric substrate in S1 is a zinc oxide substrate in the manufacturing process in this embodiment.

Example 3

The real-time ultrasound stimulation electrical signal recording chip and the manufacturing method thereof in this embodiment are the same as those in embodiment 1, except that the piezoelectric substrate in S1 is an aluminum nitride substrate in this embodiment.

Example 4

The real-time ultrasound stimulation electrical signal recording chip and the preparation method thereof in this embodiment are the same as those in embodiment 1, except that the insulating protective layer in S2 is a photoresist in the preparation of this embodiment.

Example 5

The real-time ultrasound stimulation electrical signal recording chip and the manufacturing method thereof in this embodiment are the same as those in embodiment 1, except that the insulating protective layer in S2 is polyimide in the manufacturing process in this embodiment.

Example 6

The real-time ultrasound stimulation electrical signal recording chip and the preparation method thereof in this embodiment are the same as those in embodiment 1, except that the insulating protection layer in S2 is silicon nitride during the preparation in this embodiment.

In the specific implementation, the specific processes in the above embodiments of the present invention, such as spin coating time, thickness, baking time, photoresist, developing solution, film, etc., are only used to illustrate the embodiments, and do not limit the present invention; the specific conditions of the spin coating time, the thickness and the like of the invention can be adjusted according to actual needs, other specific conditions which accord with the preparation method of the invention are all suitable for the invention, and the preparation method and/or the prepared real-time ultrasonic stimulation electric signal recording chip of the invention belong to the protection scope of the invention.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.